Compositions and methods comprising bacteria for improving behavior in neurodevelopmental disorders

ABSTRACT

Some embodiments include bacterial species for use in treatment of one or more autism spectrum disorder (ASD), and/or schizophrenia symptoms in a subject in need thereof. The bacterial species can include Bacteroides (e.g., B. fragilis, B. thetaiotaomicron, and/or B. vulgatus), and/or Enterococcus (e.g., E. faecalis, E. faecium, E. hirae, E. avium, E. durans, E. gallinarum, or E. casseliflavus). Upon treatment, one or more ASD-related behaviors can be improved in the subject.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a divisional application of U.S. applicationSer. No. 16/139,741, filed on Sep. 24, 2018, which is a continuation ofU.S. application Ser. No. 14/925,242, filed on Oct. 28, 2015, and issuedas U.S. Pat. No. 10,124,025 on Nov. 13, 2018, which claims the benefitof U.S. Provisional Application No. 62/072,873, filed on Oct. 30, 2014,each of which is hereby incorporated by reference in its entirety.

STATEMENT REGARDING FEDERALLY SPONSORED R&D

This invention was made with government support under Grant No.W81XWH-11-1-0515 awarded by the Army Research Office. The government hascertain rights in the invention.

BACKGROUND Field

Some embodiments described herein relate generally to probioticcompositions, which can be used to treat autism spectrum disorder (ASD)symptoms.

Background

Autism spectrum disorder (ASD) is a class of neurodevelopmental diseasescharacterized by the presence and severity of repetitive behaviors anddeficits in social interaction and communication. The prevalence of ASDhas continued to rise, with a current 1 in 68 children in the UnitedStates diagnosed with ASD (CDC, 2012), and similar prevalence in othercountries worldwide. ASD is believed to be caused by a combination ofgenetic and environmental risk factors. Numerous pre-clinical studiesdemonstrate that modeling maternal immune activation (MIA) in rodentsand monkeys sufficiently causes the development of ASD-relatedneuropathological and behavioral abnormalities in the offspring.

SUMMARY

In accordance with some embodiments described herein, methods forimproving a communication behavior or sensorimotor gating performance ina subject in need of such improvement are provided. In some embodiments,the method comprises identifying a subject having autism spectrumdisorder symptoms or schizophrenia symptoms in need of improvingcommunication behavior or sensorimotor gating performance. In someembodiments, the method comprises administering to the subject aneffective amount of one or more Enterococcus bacteria. In someembodiments, the subject suffers from anxiety, autism spectrum disorder(ASD), schizophrenia, or a gastrointestinal or immunological pathologyassociated with one or more of the symptoms of ASD. In some embodiments,the subject is in need of improving sensorimotor gating. In someembodiments, the subject is in need of improving sensorimotor gating anda communication behavior. In some embodiments, the communicationbehavior in need of improvement comprises at least one of impairedsociability, impaired language comprehension, impaired languageproduction, or impaired communication. In some embodiments, a soleactive ingredient administered to the subject in the method consistsessentially of the one or more Enterococcus bacteria. In someembodiments, the effective amount of the one or more Enterococcusbacteria is in a composition substantially free of bacteria other thanthe Enterococcus bacteria. In some embodiments, the one or moreEnterococcus bacteria comprises E. faecalis. In some embodiments, a soleactive ingredient administered to the subject in the method consistsessentially of E. faecalis. In some embodiments, the effective amount ofthe one or more Enterococcus bacteria is in a composition substantiallyfree of bacteria other than the E. faecalis. In some embodiments, themethod further comprises administering an effective amount of one ormore Bacteroides bacteria to the subject. In some embodiments, the oneor more Bacteroides bacteria comprises B. thetaiotaomicron, B. vulgatus,or a mixture of these bacteria. In some embodiments, the one or moreBacteroides bacteria comprises B. fragilis, B. thetaiotaomicron, B.vulgatus, or a mixture of these bacteria. In some embodiments, the oneor more Bacteroides bacteria comprises B. thetaiotaomicron, B. vulgatus,or B. fragilis or a mixture of two or three of these two bacteria (e.g.,B. thetaiotaomicron and B. vulgatus; B. thetaiotaomicron and B.fragilis, B. vulgatus, and B. fragilis, or B. thetaiotaomicron, B.vulgatus, and B. fragilis). In some embodiments, a sole activeingredient administered to the subject in the method consistsessentially of a mixture of one or more Enterococcus bacteria and one ormore Bacteroides bacteria. In some embodiments, effective amount of oneor more Enterococcus bacteria and the effective amount of one or moreBacteroides bacteria are in a composition substantially free of bacteriaother than the Enterococcus bacteria and Bacteroides bacteria. In someembodiments, a sole active ingredient administered to the subject in themethod consists essentially of: a mixture of Enterococcus bacteria andB. thetaiotaomicron; a mixture of Enterococcus bacteria and B. vulgatus;or a mixture of Enterococcus bacteria, B. thetaiotaomicron, and B.vulgatus. In some embodiments, a sole active ingredient administered tothe subject in the method consists essentially of a mixture ofEnterococcus bacteria and B. fragilis; a mixture of Enterococcusbacteria and B. thetaiotaomicron; a mixture of Enterococcus bacteria andB. vulgatus; a mixture of Enterococcus bacteria, B. fragilis, and B.thetaiotaomicron; a mixture of Enterococcus bacteria, B. fragilis, andB. vulgatus; a mixture of Enterococcus bacteria, B. thetaiotaomicron,and B. vulgatus; or a mixture of Enterococcus bacteria, B. fragilis, B.thetaiotaomicron, and B. vulgatus. In some embodiments, the effectiveamount of the one or more Enterococcus bacteria is administeredseparately from the effective amount of the one or more Bacteroidesbacteria, and each of the effective amount of the one or moreEnterococcus bacteria and the effective amount of the one or moreBacteroides bacteria are administered substantially free of bacteriaother than the Enterococcus bacteria and Bacteroides bacteria. In someembodiments, the effective amount of the one or more Enterococcusbacteria and the effective amount of the one or more Bacteroidesbacteria are in a composition substantially free of bacteria other thanEnterococcus bacteria, B. thetaiotaomicron, and B. vulgatus. In someembodiments, the effective amount of the one or more Enterococcusbacteria and the effective amount of the one or more Bacteroidesbacteria are in a composition substantially free of bacteria other thanEnterococcus bacteria, B/fragilis, B. thetaiotaomicron, and B. vulgatus.In some embodiments, the effective amount of the one or moreEnterococcus bacteria is effective when administered in combination witha Bacteroides bacteria. In some embodiments, the effective amount of oneor more Enterococcus bacteria comprises at least about 10⁷ colonyforming units (cfu), for example at least about 10⁷ cfu, at least about10⁸ cfu, at least about 10⁹ cfu, at least about 10¹⁰ cfu, at least about10¹¹ cfu, or at least about 10¹² cfu.

In some embodiments, a composition is provided. The composition cancomprise an Enterococcus bacteria and a Bacteroides bacteria. In someembodiments, the composition is substantially free of bacteria otherthan E. faecalis, B. thetaiotaomicron and B. vulgatus. In someembodiments, the composition is substantially free of bacteria otherthan E. faecalis, B. fragilis, B. thetaiotaomicron and B. vulgatus. Insome embodiments, the composition comprises a mixture of Enterococcusbacteria and B. thetaiotaomicron; a mixture of Enterococcus bacteria andB. vulgatus; or a mixture of Enterococcus bacteria, B. thetaiotaomicron,and B. vulgatus. In some embodiments, the composition comprises amixture of Enterococcus bacteria and B. thetaiotaomicron; a mixture ofEnterococcus bacteria and B. fragilis; a mixture of Enterococcusbacteria and B. vulgatus; a mixture of Enterococcus bacteria, B.thetaiotaomicron, and B. vulgatus; a mixture of Enterococcus bacteria,B. fragilis, and B. vulgatus; a mixture of Enterococcus bacteria, B.thetaiotaomicron, and B. fragilis; or a mixture of Enterococcusbacteria, B. fragilis, B. thetaiotaomicron, and B. vulgatus. In someembodiments, the composition comprises at least about 10⁷ colony formingunits (cfu) of Enterococcus bacteria, for example at least about 10⁷cfu, at least about 10⁸ cfu, at least about 10⁹ cfu, at least about 10¹⁰cfu, at least about 10¹¹ cfu, or at least about 10¹² cfu. In someembodiments, the composition comprises at least about 10⁷ colony formingunits (cfu) of Bacteroides bacteria, for example at least about 10⁷ cfu,at least about 10⁸ cfu, at least about 10⁹ cfu, at least about 10¹⁰ cfu,at least about 10¹¹ cfu, or at least about 10¹² cfu.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph illustrating that acute postnatal treatment with B.fragilis, mutant B. fragilis lacking polysaccharide A (dPSA), B.thetaiotaomicron, and B. vulgatus, but not E. faecalis corrects defectsin intestinal barrier integrity in the maternal immune activation (MIA)mouse model for autism in accordance with some embodiments describedherein.

FIGS. 2A-2B are a series of graphs illustrating that acute postnataltreatment with B. fragilis, mutant B. fragilis lacking polysaccharide A(dPSA), B. thetaiotaomicron, and B. vulgatus, but not E. faecaliscorrects anxiety-like behaviors in the maternal immune activation (MIA)mouse model for autism in accordance with some embodiments describedherein. FIG. 2A depicts center entries. FIG. 2B depicts centerduration(s).

FIG. 3 is a graph illustrating that acute postnatal treatment with B.fragilis, mutant B. fragilis lacking polysaccharide A (dPSA), B.thetaiotaomicron, and B. vulgatus, but not E. faecalis improvesrepetitive behaviors in the maternal immune activation (MIA) mouse modelfor autism in accordance with some embodiments described herein.

FIGS. 4A-4C are a series of graphs illustrating that acute postnataltreatment with B. fragilis, B. thetaiotaomicron, B. vulgatus, and E.faecalis improves communication behavior in the maternal immuneactivation (MIA) mouse model for autism in accordance with someembodiments described herein. FIG. 4A is a graph illustrating thattreatment in accordance with some embodiments described hereinsignificantly increases total number of calls produced. FIG. 4B is agraph illustrating that treatment in accordance with some embodimentsdescribed herein significantly increases average duration per call. FIG.4C is a graph illustrating that treatment in accordance with someembodiments described herein significantly increases total callduration.

FIG. 5 is a graph illustrating that acute postnatal treatment with B.fragilis, B. thetaiotaomicron, and E. faecalis improves sensorimotorgating behavior in the maternal immune activation (MIA) mouse model forautism in accordance with some embodiments described herein.

With reference to FIGS. 2A, 2B, 3, 4A, 4B, 4C, and 5, data are shown forSaline (10), Saline+B. fragilis (11), Poly(I:C) (12), Poly(I:C)+B.fragilis (13), Poly(I:C)+B. fragilis dPSA (14), Poly(I:C)+B.thetaiotaomicron (15), Poly(I:C)+B. vulgatus (16), and Poly (I:C)+E.faecilis (17).

DETAILED DESCRIPTION

In the following detailed description, reference is made to theaccompanying drawings, which form a part hereof. In the drawings,similar symbols typically identify similar components, unless contextdictates otherwise. The illustrative embodiments described in thedetailed description, drawings, and claims are not meant to be limiting.Other embodiments may be utilized, and other changes may be made,without departing from the spirit or scope of the subject matterpresented herein. It will be readily understood that the aspects of thepresent disclosure, as generally described herein, and illustrated inthe Figures, can be arranged, substituted, combined, separated, anddesigned in a wide variety of different configurations, all of which areexplicitly contemplated herein.

Without being limited by any theory, it is contemplated that there canbe a gut-immune-brain connection in autism spectrum disorders (ASD). Inaccordance with some embodiments described herein, compositionscomprising one or more bacterial species (for example, probioticcompositions) are provided, which can be administered to a subject totreat one or more symptoms of ASD, for example sensorimotor gatingbehavior deficiencies and/or communication behavior deficiencies. Insome embodiments, one or more of the following bacteria is administeredto a subject: Bacteroides fragilis, Bacteroides thetaiotaomicron,Bacteroides vulgatus, and Enterococcus faecalis, so as to improvesensorimotor gating behavior and/or communication behavior deficienciesin the subject.

Maternal infection is regarded as a primary environmental risk factorfor ASD, as well as other neurodevelopmental disorders such asschizophrenia. Several large epidemiological studies indicate thatmaternal bacterial or viral infection during pregnancy increases therisk for ASD in the offspring. Similar findings link elevated levels ofimmune signaling factors, such as cytokines and chemokines, in thematernal serum or amniotic fluid during pregnancy to increased ASD risk.

In addition to its core diagnostic features, ASD is also associated withseveral co-morbid medical conditions. Immune dysregulation andgastrointestinal issues are of particular interest, in light of theirhigh prevalence in ASD and their correlation with the severity ofcardinal ASD-related behavioral impairments. A significant subset ofindividuals with ASD present with gastrointestinal distress, includingconstipation, abdominal pain, immune activation, and intestinal barrierdysfunction (“leaky gut”). In addition, several studies report that thecomposition of gut bacteria (the intestinal microbiome) is altered inchildren with ASD compared to controls. Without being limited by anytheory, it is contemplated herein that there is a potentialgut-immune-brain connection in ASD.

Definitions

Unless defined otherwise, technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which the present disclosure belongs. See, e.g. Singleton etal., Dictionary of Microbiology and Molecular Biology 2nd ed., J. Wiley& Sons (New York, N.Y. 1994); Sambrook et al., Molecular Cloning, ALaboratory Manual, Cold Springs Harbor Press (Cold Springs Harbor, N.Y.1989). For purposes of the present disclosure, the following terms aredefined below.

As used herein, the term “subject” is a vertebrate, such as a mammal.The term “mammal” is defined as an individual belonging to the classMammalia and includes, without limitation, humans, domestic and farmanimals, and zoo, sports, or pet animals, such as sheep, dogs, horses,cats, or cows. In some embodiments, the subject is human. In someembodiments, the subject is a non-human primate.

As used herein, the term “condition/disorder/symptom” or “behavioralabnormality” refers to a symptom expressed by a subject including butnot limited to anxiety, Fragile X, Rett syndrome, tuberous sclerosis,obsessive compulsive disorder, attention deficit disorder,schizophrenia, autistic disorder (classic autism), Asperger's disorder(Asperger syndrome), pervasive developmental disorder not otherwisespecified (PDD-NOS), childhood disintegrative disorder (CDD), or apathological condition with one or more of the symptoms of ASD.

As used herein, the term “subject in need of the treatment” refers to asubject expressing or suffering from one or more of the behavioraldisorder/symptoms mentioned above. In some embodiments, the subject inneed of treatment suffers from at least one of schizophrenia, ASD, or agastrointestinal or immunological pathology associated with ASD orschizophrenia (for example leaky gut syndrome). An appropriatelyqualified person is able to identify such an individual in need oftreatment using standard behavioral testing protocols/guidelines. Thesame behavioral testing protocols/guidelines can also be used todetermine whether there is improvement to the individual's disorderand/or symptoms. As used herein “sensorimotor gating” refers to abilityto filter out irrelevant and/or intrusive sensory stimuli. As such,subjects deficient in sensorimotor gating behavior can have impairedability to filter out stimuli, and/or can have difficulty coping withintensely stimulating environments. By way of example, sensorimotorgating behavior can be assessed with a prepulse inhibition (PPI) test.In accordance with some embodiments described herein a subject isidentified as being in need of improved sensorimotor gating, for examplein need of improving filtering out of irrelevant sensory stimuli such asbackground noise, background light, and the like. As used herein,“communication behavior” refers to communication, language comprehensionand production, and/or sociability, including vocal and non-vocal socialcommunication. In some embodiments, a subject is identified as deficientin communication behavior based on impaired sociability. In someembodiments, a subject is identified as deficient in communicationbehavior based on impaired language comprehension and/or production.

As used herein, the term “improvement in behavioral performance” refersto prevention or reduction in the severity or frequency, to whateverextent, of one or more of the behavioral disorders, symptoms and/orabnormalities expressed by individual suffering from ASD, schizophrenia,or a pathological condition with one or more of the symptoms of ASD orschizophrenia. Non-limiting examples of the behavioral symptoms includeimpaired communication, impaired sociability, impaired languagecomprehension and/or production, impaired sensorimotor gating behavior,repetitive behavior, and increased anxiety. The improvement is eitherobserved by the individual taking the treatment themselves or by anotherperson (medical or otherwise). In some embodiments, a probioticcomprising an effective amount of Bacteroides and/or Enterococcusbacteria as described herein is administered the subject. In someembodiments, sensorimotor gating behavior is improved in the subjectafter administration of the probiotic. In some embodiments,communication behavior is improved in the subject after administrationof the probiotic. Examples of communication behaviors that can beimproved include communication, sociability, and language comprehensionand/or production. In some embodiments, anxiety behavior is improved inthe subject after administration of the probiotic. In some embodiments,repetitive behavior is improved in the subject after administration ofthe probiotic. In some embodiments, sensorimotor gating behavior andcommunication behavior are improved in the subject after administrationof the probiotic.

As used herein, the term “treatment” refers to a clinical interventionmade in response to a disease, disorder or physiological conditionmanifested by a subject, particularly a subject suffering from ASD,schizophrenia, or a pathological condition with one or more of thesymptoms of ASD or schizophrenia. The aim of treatment may include, butis not limited to, one or more of the alleviation or prevention ofsymptoms, slowing or stopping the progression or worsening of a disease,disorder, or condition and the remission of the disease, disorder, orcondition. In some embodiments, “treatment” refers to both therapeutictreatment and prophylactic or preventative measures. Those in need oftreatment include those already affected by a disease or disorder orundesired physiological condition as well as those in which the diseaseor disorder or undesired physiological condition is to be prevented. Forexample, in some embodiments, treatment may improve behavioralperformance of the subject, including ASD-related behaviors such assensorimotor gating behavior deficiencies and/or communication behaviordeficiencies. As used herein, the term “prevention” refers to anyactivity that reduces the burden of the individual later expressingthose behavioral symptoms. This takes place at primary, secondary andtertiary prevention levels, wherein: a) primary prevention avoids thedevelopment of symptoms/disorder/condition; b) secondary preventionactivities are aimed at early stages of the condition/disorder/symptomtreatment, thereby increasing opportunities for interventions to preventprogression of the condition/disorder/symptom and emergence of symptoms;and c) tertiary prevention reduces the negative impact of an alreadyestablished condition/disorder/symptom by, for example, restoringfunction and/or reducing any condition/disorder/symptom or relatedcomplications.

“Pharmaceutically acceptable” carriers are ones which are nontoxic tothe cell or mammal being exposed thereto at the dosages andconcentrations employed. “Pharmaceutically acceptable” carriers inaccordance with methods and uses and compositions and kits herein cancomprise, but not limited to, organic or inorganic, solid or liquidexcipients which is suitable for the selected mode of application suchas oral application or injection, and administered in the form of aconventional pharmaceutical preparation, such as solid such as tablets,granules, powders, capsules, and liquid such as solution, emulsion,suspension, and the like. Often the physiologically acceptable carrieris an aqueous pH buffered solution such as phosphate buffer or citratebuffer. The physiologically acceptable carrier may also comprise one ormore of the following: antioxidants including ascorbic acid, lowmolecular weight (less than about 10 residues) polypeptides, proteins,such as serum albumin, gelatin, immunoglobulins; hydrophilic polymerssuch as polyvinylpyrrolidone, amino acids, carbohydrates includingglucose, mannose, or dextrins, chelating agents such as EDTA, sugaralcohols such as mannitol or sorbitol, salt-forming counterions such assodium, and nonionic surfactants such as TWEEN™ surfactant, polyethyleneglycol (PEG), and PLURONICS™ surfactant. Auxiliary, stabilizer,emulsifier, lubricant, binder, pH adjustor controller, isotonic agentand other conventional additives may also be added to the carriers.

The pharmaceutically acceptable or appropriate carrier in accordancewith methods and uses and compositions and kits herein may include othercompounds known to be beneficial to an impaired situation of the GItract (e.g., antioxidants, such as Vitamin C, Vitamin E, Selenium orZinc); or a food composition. The food composition can be, but is notlimited to, milk, yoghurt, curd, cheese, fermented milks, milk basedfermented products, ice-creams, fermented cereal-based products,milk-based powders, infant formulae, tablets, liquid bacterialsuspensions, dried oral supplement, or wet oral supplement.

As used herein, the term “probiotic” refers to live microorganisms,which, when administered in adequate amounts, confer a health benefit onthe host. The probiotics in accordance with methods and uses andcompositions and kits herein may be available in foods and dietarysupplements (for example, but not limited to capsules, tablets, powders,and liquids). Non-limiting examples of foods containing probioticinclude dairy products such as yogurt, fermented and unfermented milk,smoothies, butter, cream, hummus, kombucha, salad dressing, miso,tempeh, nutrition bars, and some juices and soy beverages. In someembodiments, the probiotic comprises a single microorganism. In someembodiments, the probiotic comprises a combination of microorganisms. Insome embodiments, the probiotic comprises a single composition. In someembodiments, the probiotic comprises two or more compositions, which canbe used together, for example administered simultaneously oradministered sequentially. It is noted that a probiotic can serve as the“active ingredient” or a composition or compositions for use inadministration to a subject. That is, the method, use, and/orcomposition or compositions (either individually or in the aggregate)can comprise an effective amount of probiotic to improve at least onebehavior in a subject. In some embodiments, the probiotic is the soleactive ingredient for administration to the subject. In someembodiments, the “sole active ingredient” probiotic for administrationto the subject can be provided in a composition or in a method or usethat is substantially free of or free of bacteria other than theprobiotic, antibiotics, and drugs. Even if the probiotic is the “sole”active ingredient, the composition or composition comprising theprobiotic may comprise additional substances (such as buffers, bacterialfeedstock, excipients, flavors, and/or food) that do not substantiallyaffect the behavior of the subject, but may be useful for the functionof the probiotic or its administration.

In some embodiments, the probiotic is comprised in a composition orcompositions that are substantially free of bacteria (other than theprobiotic) and/or drugs or antibiotics. By “substantially free” or“substantially absent”, it is understood that while a bacteria otherthan the probiotic, drug, and/or antibiotic may be present in traceamounts, the bacteria other than the probiotic, drug, and/or antibiotichave no appreciable effect on the subject.

As used herein “effective amount” of probiotic refers to a quantitysufficient to achieve a clinically significant change in a behavior of asubject.

As used herein, the term “nutraceutical” refers to a food stuff (as afortified food or a dietary supplement) that provides health benefits.Nutraceutical foods are not subject to the same testing and regulationsas pharmaceutical drugs.

Probiotics for Treatment of ASD and/or Schizophrenia Symptoms

Without being limited by any theory, it is contemplated that the MIAmouse model for autism, which displays both neuropathological andbehavioral features of ASD and also schizophrenia, and also exhibitsimmunological and gastrointestinal abnormalities relevant to the humandisorder. It is demonstrated herein that treatment of MIA offspring withhuman commensal bacteria such as Bacteroides fragilis correctsparticular gastrointestinal and behavioral deficits. Accordingly, someembodiments include a probiotic treatment for symptoms of ASD and/orschizophrenia.

Bacteroides fragilis has been observed to have immunomodulatoryproperties. Unexpectedly, it has been demonstrated herein that otherbacterial species can confer a probiotic treatment in MIA mice. 3-weekold MIA offspring were treated with Bacteroides thetaiotaomicron,Bacteroides vulgatus, or Enterococcus faecalis for one week, and thenassayed for ASD-related gastrointestinal and behavioral symptoms. It isshown that treatment with B. thetaiotaomicron or B. vulgatus correctsdeficits in intestinal barrier integrity in MIA offspring, whereastreatment with E. faecilis has no significant effect (see FIG. 1). Inaddition, B. thetaiotaomicron and B. vulgatus treatment improveanxiety-like, repetitive and communication behavior in MIA mice (seeFIGS. 2-4). B. thetaiotaomicron and E. faecalis improve sensorimotorgating behavior deficiencies in MIA mice (FIG. 5). Accordingly, in someembodiments described herein, probiotic treatments for ASD-relatedbehaviors are provided. The treatment can comprise administering aprobiotic comprising, consisting essentially of, or consisting of B.fragilis, B. thetaiotaomicron, B. vulgatus, or E. faecilis, or acombination of two, three, or four of these bacteria as described hereinto a subject in need of behavioral performance. The subject can be inneed of improvement in sensorimotor gating, and/or communicationbehavior.

In some embodiments, the subject is in need of improvement insensorimotor gating behavior, and an effective amount of a probioticcomprising, consisting of, or consisting essentially of at least one ofthe following is provided for administration to the subject (or is foruse in treating the subject): (a) Enterococcus bacteria (e.g., E.faecilis E. faecium, E. hirae, E. avium, E. durans, E. gallinarum, or E.casseliflavus); (b) Enterococcus bacteria (e.g., E. faecilis, E.faecium, E. hirae, E. avium, E. durans, E. gallinarum, or E.casseliflavus); and Bacteroides bacteria (e.g., B. fragilis, B.thetaiotaomicron or B. vulgatus); (c) Enterococcus bacteria and B.fragilis; (d) Enterococcus bacteria and B. thetaiotaomicron; (d)Enterococcus bacteria and B. vulgatus; (e) Enterococcus bacteria, B.fragilis, and B. thetaiotaomicron, (f) Enterococcus bacteria, B.fragilis, and B. vulgatus; (g) Enterococcus bacteria, B.thetaiotaomicron and B. vulgatus; (h) Enterococcus bacteria, B.fragilis, B. thetaiotaomicron and B. vulgatus; (i) E. faecilis and B.fragilis; (j) E. faecilis and B. thetaiotaomicron; (k) E. faecilis andB. vulgatus; (l) E. faecilis, B. fragilis, and B. thetaiotaomicron, (m)E. faecilis, B. fragilis, and B. vulgatus; (n) E. faecilis, B.thetaiotaomicron and B. vulgatus; (o) E. faecilis, B. fragilis, B.thetaiotaomicron and B. vulgatus; (p) Bacteroides bacteria; (q) B.fragilis and B. thetaiotaomicron; (r) B. fragilis, and B. vulgatus; (s)B. thetaiotaomicron and B. vulgatus; or (t) B. fragilis, B.thetaiotaomicron and B. vulgatus. Following administration of thebacteria, the sensorimotor gating behavior can be improved. Optionally,the subject is administered no other bacteria, or substantially no otherbacteria apart from the identified bacteria of the probiotic, and assuch the probiotic for use in treatment of the subject is in acomposition or compositions free or substantially free of otherbacteria. Optionally, the subject is administered no antibiotics, or isadministered substantially no antibiotics, and as such the probiotic foradministration to the subject is in a composition or compositions freeor substantially free of antibiotics. Optionally, the subject isadministered no drugs, or is administered substantially no drugs, and assuch the probiotic for administration to the subject is in a compositionor compositions free or substantially free of drugs. Optionally, thesubject is administered no pharmaceutically active ingredients, or isadministered substantially no pharmaceutically active ingredients, andas such the probiotic for administration to the subject is in acomposition or compositions free or substantially free ofpharmaceutically active ingredients. Optionally, the B. fragiliscomprises wild-type B. fragilis, mutant B. fragilis lackingpolysaccharide A (dPSA), or a combination of wild-type B. fragilis anddPSA B. fragilis. In some embodiments, the sensorimotor gating behaviorcomprises or correlates with frequency of pre-pulse inhibition. In someembodiments, the subject has communication behavior deficienciesassociated with ASD, for example deficient communication, sociability,and language comprehension and/or production. In some embodiments, thesubject has sensorimotor gating behavior deficiencies associated withASD. In some embodiments, the subject has communication behaviordeficiencies associated with schizophrenia, for example deficientcommunication, sociability, and language comprehension and/orproduction. In some embodiments, the subject has sensorimotor gatingbehavior deficiencies associated with schizophrenia.

In some embodiments, the subject is in need of improvement incommunication behavior, and an effective amount of a probioticcomprising, consisting of, or consisting essentially of at least one ofthe following is provided for administration to the subject (or is foruse in treating the subject): (a) Enterococcus bacteria (e.g., E.faecilis, E. faecium, E. hirae, E. avium, E. durans, E. gallinarum, orE. cassehflavus); (b) Enterococcus bacteria (e.g., E. faecilis, E.faecium, E. hirae, E. avium, E. durans, E. galhnarum, or E.cassehflavus); and Bacteroides bacteria (e.g., B. fragilis, B.thetaiotaomicron or B. vulgatus); (c) Enterococcus bacteria and B.fragilis, (d) Enterococcus bacteria and B. thetaiotaomicron; (d)Enterococcus bacteria and B. vulgatus; (e) Enterococcus bacteria, B.fragilis, and B. thetaiotaomicron, (f) Enterococcus bacteria, B.fragilis, and B. vulgatus; (g) Enterococcus bacteria, B.thetaiotaomicron and B. vulgatus; (h) Enterococcus bacteria, B.fragilis, B. thetaiotaomicron and B. vulgatus; (i) E. faecilis and B.fragilis; (j) E. faecilis and B. thetaiotaomicron; (k) E. faecilis andB. vulgatus; (l) E. faecilis, B. fragilis, and B. thetaiotaomicron, (m)E. faecilis, B. fragilis, and B. vulgatus; (n) E. faecilis, B.thetaiotaomicron and B. vulgatus; (o) E. faecilis, B. fragilis, B.thetaiotaomicron and B. vulgatus; (p) Bacteroides bacteria; (q) B.fragilis and B. thetaiotaomicron; (r) B. fragilis, and B. vulgatus; (s)B. thetaiotaomicron and B. vulgatus; or (t) B. fragilis, B.thetaiotaomicron and B. vulgatus. Following administration of thebacteria, the vocalization and/or communication behavior can beimproved. In some embodiments, the communication behavior in need ofimprovement (and subsequently improved) comprises at least one ofcommunication, sociability, or language comprehension and/or production.Optionally, the subject is administered no other bacteria, orsubstantially no other bacteria apart from the identified bacteria ofthe probiotic, and as such the probiotic for use in treatment of thesubject is in a composition or compositions free or substantially freeof other bacteria. Optionally, the subject is administered noantibiotics, or is administered substantially no antibiotics, and assuch the probiotic for administration to the subject is in a compositionor compositions free or substantially free of antibiotics. Optionally,the subject is administered no drugs, or is administered substantiallyno drugs, and as such the probiotic for administration to the subject isin a composition or compositions free or substantially free of drugs.Optionally, the subject is administered no pharmaceutically activeingredients, or is administered substantially no pharmaceutically activeingredients, and as such the probiotic for administration to the subjectis in a composition or compositions free or substantially free ofpharmaceutically active ingredients. Optionally, the B. fragiliscomprises wild-type B. fragilis, mutant B. fragilis lackingpolysaccharide A (dPSA), or a combination of wild-type B. fragilis anddPSA B. fragilis. In some embodiments, the subject has communicationbehavior deficiencies associated with ASD, for example deficientcommunication, sociability, and language comprehension and/orproduction. In some embodiments, the subject has sensorimotor gatingbehavior deficiencies associated with ASD. In some embodiments, thesubject has communication behavior deficiencies associated withschizophrenia, for example deficient communication, sociability, andlanguage comprehension and/or production. In some embodiments, thesubject has sensorimotor gating behavior deficiencies associated withschizophrenia.

In some embodiments, the subject is in need of improvement insensorimotor gating and/or communication behavior, and an effectiveamount of a probiotic comprising, consisting of, or consistingessentially of at least one of the following is provided foradministration to the subject (or is for use in treating the subject):(a) Enterococcus bacteria (e.g., E. faecilis, E. faecium, E. hirae, E.avium, E. durans, E. gallinarum, or E. casseliflavus); (b) Enterococcusbacteria (e.g., E. faecilis, E. faecium, E. hirae, E. avium, E. durans,E. gallinarum, or E. casseliflavus); and Bacteroides bacteria (e.g., B.fragilis, B. thetaiotaomicron or B. vulgatus); (c) Enterococcus bacteriaand B. fragilis; (d) Enterococcus bacteria and B. thetaiotaomicron; (d)Enterococcus bacteria and B. vulgatus; (e) Enterococcus bacteria, B.fragilis, and B. thetaiotaomicron, (f) Enterococcus bacteria, B.fragilis, and B. vulgatus; (g) Enterococcus bacteria, B.thetaiotaomicron and B. vulgatus; (h) Enterococcus bacteria, B.fragilis, B. thetaiotaomicron and B. vulgatus; (i) E. faecilis and B.fragilis; (j) E. faecilis and B. thetaiotaomicron; (k) E. faecilis andB. vulgatus; (l) E. faecilis, B. fragilis, and B. thetaiotaomicron, (m)E. faecilis, B. fragilis, and B. vulgatus; (n) E. faecilis, B.thetaiotaomicron and B. vulgatus; (o) E. faecilis, B. fragilis, B.thetaiotaomicron and B. vulgatus; (p) Bacteroides bacteria; (q) B.fragilis and B. thetaiotaomicron; (r) B. fragilis, and B. vulgatus; (s)B. thetaiotaomicron and B. vulgatus; or (t) B. fragilis, B.thetaiotaomicron and B. vulgatus. Following administration of thebacteria, the sensorimotor gating and/or communication behavior can beimproved. In some embodiments, the subject is in need of improvedsensorimotor gating behavior. In some embodiments, the subject is inneed of improved communication behavior. In some embodiments, thesubject is in need of improved sensorimotor gating and communicationbehavior. In some embodiments, the communication behavior in need ofimprovement (and subsequently improved) comprises at least one ofcommunication, sociability, or language comprehension and/or languageproduction. Optionally, the subject is administered no other bacteria,or substantially no other bacteria apart from the identified bacteria ofthe probiotic, and as such the probiotic for use in treatment of thesubject is in a composition or compositions free or substantially freeof other bacteria. Optionally, the subject is administered noantibiotics, or is administered substantially no antibiotics, and assuch the probiotic for administration to the subject is in a compositionor compositions free or substantially free of antibiotics. Optionally,the subject is administered no drugs, or is administered substantiallyno drugs, and as such the probiotic for administration to the subject isin a composition or compositions free or substantially free of drugs.Optionally, the subject is administered no pharmaceutically activeingredients, or is administered substantially no pharmaceutically activeingredients, and as such the probiotic for administration to the subjectis in a composition or compositions free or substantially free ofpharmaceutically active ingredients. Optionally, the B. fragiliscomprises wild-type B. fragilis, mutant B. fragilis lackingpolysaccharide A (dPSA), or a combination of wild-type B. fragilis anddPSA B. fragilis. In some embodiments, the subject is further in need ofimprovement of defects in intestinal barrier integrity, and followingadministration of the probiotic, the defects in defects in intestinalbarrier integrity are improved. In some embodiments, the subject hascommunication behavior deficiencies associated with ASD, for exampledeficient communication, sociability, and language comprehension and/orproduction. In some embodiments, the subject has sensorimotor gatingbehavior deficiencies associated with ASD. In some embodiments, thesubject has communication behavior deficiencies associated withschizophrenia, for example deficient communication, sociability, andlanguage comprehension and/or production. In some embodiments, thesubject has sensorimotor gating behavior deficiencies associated withschizophrenia. In some embodiments, the subject has ASD. In someembodiments, the subject has schizophrenia.

In some embodiments, the subject is in need of improvement in deficitsin intestinal barrier integrity, and an effective amount of a probioticcomprising, consisting of, or consisting essentially of at least one ofthe following is provided for administration to the subject (or is foruse in treating the subject): (a) Bacteroides bacteria; (b) B. fragilisand B. thetaiotaomicron; (c) B. fragilis, and B. vulgatus; (d) B.thetaiotaomicron and B. vulgatus; or (e) B. fragilis, B.thetaiotaomicron and B. vulgatus. Following administration of thebacteria, the deficits in intestinal barrier integrity can be improved.Optionally, the subject is administered no other bacteria, orsubstantially no other bacteria apart from the identified bacteria ofthe probiotic, and as such the probiotic for use in treatment of thesubject is in a composition or compositions free or substantially freeof other bacteria. Optionally, the subject is administered noantibiotics, or is administered substantially no antibiotics, and assuch the probiotic for administration to the subject is in a compositionor compositions free or substantially free of antibiotics. Optionally,the subject is administered no drugs, or is administered substantiallyno drugs, and as such the probiotic for administration to the subject isin a composition or compositions free or substantially free of drugs.Optionally, the subject is administered no pharmaceutically activeingredients, or is administered substantially no pharmaceutically activeingredients, and as such the probiotic for administration to the subjectis in a composition or compositions free or substantially free ofpharmaceutically active ingredients. Optionally, the B. fragiliscomprises wild-type B. fragilis, mutant B. fragilis lackingpolysaccharide A (dPSA), or a combination of wild-type B. fragilis anddPSA B. fragilis. In some embodiments, the subject has communicationbehavior deficiencies associated with ASD, for example deficientcommunication, sociability, and language comprehension and/orproduction. In some embodiments, the subject has sensorimotor gatingbehavior deficiencies associated with ASD. In some embodiments, thesubject has communication behavior deficiencies associated withschizophrenia, for example deficient communication, sociability, andlanguage comprehension and/or production. In some embodiments, thesubject has sensorimotor gating behavior deficiencies associated withschizophrenia. In some embodiments, the subject has defects inintestinal barrier integrity associated with ASD. In some embodiments,the subject has defects in intestinal barrier integrity associated withschizophrenia. In some embodiments, the subject has ASD. In someembodiments, the subject has schizophrenia.

In some embodiments, the subject is in need of improvement in anxietybehavior, and an effective amount of a probiotic comprising, consistingof, or consisting essentially of at least one of the following isprovided for administration to the subject (or is for use in treatingthe subject): (a) Bacteroides bacteria; (b) B. fragilis and B.thetaiotaomicron; (c) B. fragilis, and B. vulgatus; (d) B.thetaiotaomicron and B. vulgatus; or (e) B. fragilis, B.thetaiotaomicron and B. vulgatus. Following administration of thebacteria, the anxiety behavior can be improved. Optionally, the subjectis administered no other bacteria, or substantially no other bacteriaapart from the identified bacteria of the probiotic, and as such theprobiotic for use in treatment of the subject is in a composition orcompositions free or substantially free of other bacteria. Optionally,the subject is administered no antibiotics, or is administeredsubstantially no antibiotics, and as such the probiotic foradministration to the subject is in a composition or compositions freeor substantially free of antibiotics. Optionally, the subject isadministered no drugs, or is administered substantially no drugs, and assuch the probiotic for administration to the subject is in a compositionor compositions free or substantially free of drugs. Optionally, thesubject is administered no pharmaceutically active ingredients, or isadministered substantially no pharmaceutically active ingredients, andas such the probiotic for administration to the subject is in acomposition or compositions free or substantially free ofpharmaceutically active ingredients. Optionally, the B. fragiliscomprises wild-type B. fragilis, mutant B. fragilis lackingpolysaccharide A (dPSA), or a combination of wild-type B. fragilis anddPSA B. fragilis. In some embodiments, the subject is further in need ofimprovement of defects in intestinal barrier integrity, and followingadministration of the probiotic, the defects in defects in intestinalbarrier integrity are improved. In some embodiments, the anxietybehavior is associated with ASD or schizophrenia. In some embodiments,the subject has ASD. In some embodiments, the subject has schizophrenia.

In some embodiments, the subject is in need of improvement in repetitivebehavior, and an effective amount of a probiotic comprising, consistingof, or consisting essentially of at least one of the following isprovided for administration to the subject (or is for use in treatingthe subject): (a) Bacteroides bacteria; (b) B. fragilis and B.thetaiotaomicron; (c) B. fragilis, and B. vulgatus; (d) B.thetaiotaomicron and B. vulgatus; or (e) B. fragilis, B.thetaiotaomicron and B. vulgatus. Following administration of thebacteria, the repetitive behavior can be improved. Optionally, thesubject is administered no other bacteria, or substantially no otherbacteria apart from the identified bacteria of the probiotic, and assuch the probiotic for use in treatment of the subject is in acomposition or compositions free or substantially free of otherbacteria. Optionally, the subject is administered no antibiotics, or isadministered substantially no antibiotics, and as such the probiotic foradministration to the subject is in a composition or compositions freeor substantially free of antibiotics. Optionally, the subject isadministered no drugs, or is administered substantially no drugs, and assuch the probiotic for administration to the subject is in a compositionor compositions free or substantially free of drugs. Optionally, thesubject is administered no pharmaceutically active ingredients, or isadministered substantially no pharmaceutically active ingredients, andas such the probiotic for administration to the subject is in acomposition or compositions free or substantially free ofpharmaceutically active ingredients. Optionally, the B. fragiliscomprises wild-type B. fragilis, mutant B. fragilis lackingpolysaccharide A (dPSA), or a combination of wild-type B. fragilis anddPSA B. fragilis. In some embodiments, the subject is further in need ofimprovement of defects in intestinal barrier integrity, and followingadministration of the probiotic, the defects in defects in intestinalbarrier integrity are improved. In some embodiments, the repetitivebehavior is associated with ASD or schizophrenia. In some embodiments,the subject has ASD. In some embodiments, the subject has schizophrenia.

In some embodiments, the probiotic comprises any of the above-disclosedbacterial species or combinations of bacterial species, and is providedfor administration to the subject (or is for administration to thesubject) in a single probiotic composition. In some embodiments, theprobiotic comprises any of the above-referenced bacterial species orcombinations of bacterial species, and is administered to the subject(or is for administration to the subject) in two or more differentprobiotic compositions. For example, a probiotic of “bacteria A andbacteria B” can be administered either in a single compositioncomprising bacteria A and bacteria B, or in a first compositioncomprising bacteria A in conjunction with a second compositioncomprising bacteria B. In some embodiments, first and secondcompositions are administered simultaneously. In some embodiments, thefirst and second compositions are administered separately.

In some embodiments, a probiotic comprising a combination ofEnterococcus bacteria and Bacteroides bacteria as described herein isprovided as a first composition comprising the Enterococcus bacteria,and a second composition comprising the Bacteroides bacteria orcombination of Bacteroides bacteria as described herein. In someembodiments, the Enterococcus bacteria and at least one Bacteroides (orcombination of Bacteroides bacteria) are administered in a firstcomposition, and at least one different Bacteroides bacteria (ordifferent combination of Bacteroides bacteria) is administered in asecond composition. In some embodiments, the Enterococcus bacteria and afirst Bacteroides bacteria (or combination of Bacteroides bacteria) isadministered in a first composition, and the Enterococcus bacteria and asecond Bacteroides bacteria (or combination of Bacteroides bacteria)that is different from the first is administered in a secondcomposition. In some embodiments, the Enterococcus bacteria and a firstBacteroides bacteria (or combination of Bacteroides bacteria) isadministered in a first composition, and a second Bacteroides bacteria(or combination of Bacteroides bacteria) that is different from thefirst is administered in a second composition.

In some embodiments, a probiotic comprising a combination of Bacteroidesbacteria as described herein is administered via a first compositioncomprising a first Bacteroides bacteria or combination of Bacteroidesbacteria, and a second composition comprising a second Bacteroidesbacteria or combination of Bacteroides bacteria that is different fromthe first.

In accordance with any of the embodiments described above, optionally,each composition, use or method is free of, or is substantially free ofbacteria other than the identified bacteria of the probiotic. Inaccordance with any of the embodiments above, optionally, eachcomposition is free of, or is substantially free of antibiotics. Inaccordance with any of the embodiments above, optionally, eachcomposition is free of, or is substantially free of bacteria other thanthe probiotic and antibiotics.

In accordance with embodiments described herein, the probiotics of themethods, uses, and compositions described herein can be for any suitableroute of administration. For example, the probiotic can be administeredto the subject via oral administration, rectum administration,transdermal administration, intranasal administration, or inhalation. Insome embodiments, the probiotic is administered to the subject orally.

In some embodiments, the effective amount of bacteria in the probioticcomposition, use, or method includes at least about 10⁴ colony formingunits (cfu), for example at least about 10⁴, 10⁵, 10⁶, 10⁷, 10⁸, 10⁹,10¹⁰, 10¹¹, 10¹², or 10¹³ cfu, including ranges between any of thelisted values, for example 10⁴-10⁸ cfu, 10⁴-10⁹ cfu, 10⁴-10¹⁰ cfu,10⁴-10¹¹ cfu, 10⁴-10¹² cfu, 10⁵-10⁸ cfu, 10⁵-10⁹ cfu, 10⁵-10¹⁰ cfu,10⁵-10¹¹ cfu, 10⁵-10¹² cfu, 10⁶-10⁸ cfu, 10⁶-10⁹ cfu, 10⁶-10¹⁰ cfu,10⁶-10¹¹ cfu, 10⁶-10¹² cfu, 10⁷-10⁸ cfu, 10⁷-10⁹ cfu, 10⁷-10¹⁰ cfu,10⁷-10¹¹ cfu, 10⁷-10¹² cfu, 10⁸-10⁹ cfu, 10⁸-10¹⁰ cfu, 10⁸-10¹¹ cfu, or10⁸-10¹² cfu. In some embodiments, the effective amount of bacteriacomprises a log phase quantity (at 37° C.) of bacteria in a compositionfor administration to the subject. In some embodiments, the effectiveamount of bacteria comprises a stationary phase quantity (at 37° C.) ofbacteria in a composition for administration to the subject.

Methods of Treating and/or Preventing ASD and/or Schizophrenia Symptoms

In some embodiments, methods of treating ASD and/or schizophreniasymptoms are provided. The method can comprise identifying a subject asin need of improving a sensorimotor gating behavior, and/orcommunication behavior. The method can comprise administering aneffective amount of a probiotic comprising, consisting essentially of,or consisting of Enterococcus bacteria, Bacteroides bacteria, or acombination of Enterococcus bacteria and Bacteroides bacteria asdescribed herein is administered to the subject in need of improvedsensorimotor gating behavior and/or communication behavior. The subjectcan exhibit improved sensorimotor gating behavior and/or communicationbehavior. In some embodiments, the subject is in need of improvedsensorimotor gating behavior, and following administration of theprobiotic, sensorimotor gating behavior is improved. In someembodiments, the subject is in need of improved communication behavior,and following administration of the probiotic, communication behavior isimproved. In some embodiments, the subject is in need of improvedcommunication and sensorimotor gating behavior, and followingadministration of the probiotic, communication behavior and sensorimotorgating behavior are improved. In some embodiments, the communicationbehavior in need of improvement (and subsequently improved) comprises atleast one of communication, sociability, or language comprehensionand/or language production. In some embodiments, the subject has ASD. Insome embodiments, the method further comprises determining whether thesubject has ASD, and the effective amount of probiotic is administeredif the subject has ASD. In some embodiments, the subject hasschizophrenia. In some embodiments, the method further comprisesdetermining whether the subject has schizophrenia, and the effectiveamount of probiotic is administered if the subject has schizophrenia.

In some embodiments, the subject is in need of improved anxietybehavior. The method can comprise identifying the subject as in need ofimproved anxiety behavior. The method can comprise administering aneffective amount of a probiotic comprising, consisting essentially of,or consisting of Enterococcus bacteria, Bacteroides bacteria, or acombination of Enterococcus bacteria and Bacteroides bacteria asdescribed herein can be administered to the subject in need of improvedanxiety behavior, and following administration of the probiotic, anxietybehavior is improved. In some embodiments, the subject is in need ofimproved sensorimotor gating behavior and anxiety behavior, andfollowing administration of the probiotic, sensorimotor gating behaviorand communication behavior are improved. In some embodiments, thesubject is in need of improved communication behavior and anxietybehavior, and following administration of the probiotic, communicationbehavior and anxiety behavior are improved. In some embodiments, thesubject is in need of improved communication behavior, and followingadministration of the probiotic, communication behavior is improved. Insome embodiments, the communication behavior in need of improvement (andsubsequently improved) comprises at least one of communication,sociability, or language comprehension and/or language production. Insome embodiments, the subject is in need of improved sensorimotor gatingbehavior, communication behavior, and anxiety behavior, and followingadministration of the probiotic, sensorimotor gating behavior andanxiety behavior are improved. In some embodiments, the subject isfurther identified as in need of improvement of defects in intestinalbarrier integrity, and following administration of the probiotic, thedefects in defects in intestinal barrier integrity are improved. In someembodiments, the subject has ASD. In some embodiments, the methodfurther comprises determining whether the subject has ASD, and theeffective amount of probiotic is administered if the subject has ASD. Insome embodiments, the subject has schizophrenia. In some embodiments,the method further comprises determining whether the subject hasschizophrenia, and the effective amount of probiotic is administered ifthe subject has schizophrenia.

In some embodiments, methods of treating ASD or schizophrenia symptomsare provided. The method can comprise identifying the subject as in needof improving sensorimotor gating behavior. The method can compriseadministering an effective amount of a probiotic comprising, consistingessentially of, or consisting of Enterococcus bacteria, Bacteroidesbacteria, or a combination of Enterococcus bacteria and Bacteroidesbacteria as described herein is administered to the subject. Followingadministration of the probiotic, sensorimotor gating behavior in thesubject can be improved. In some embodiments, the subject is furtheridentified as in need of improvement of defects in intestinal barrierintegrity, and following administration of the probiotic, the defects indefects in intestinal barrier integrity are improved. In someembodiments, the subject has ASD. In some embodiments, the methodfurther comprises determining whether the subject has ASD, and theeffective amount of probiotic is administered if the subject has ASD. Insome embodiments, the subject has schizophrenia. In some embodiments,the method further comprises determining whether the subject hasschizophrenia, and the effective amount of probiotic is administered ifthe subject has schizophrenia.

In some embodiments, methods of treating ASD or schizophrenia symptomsare provided. The method can comprise identifying the subject as in needof improving communication behavior. The method can compriseadministering an effective amount of a probiotic comprising, consistingessentially of, or consisting of Enterococcus bacteria, Bacteroidesbacteria, or a combination of Enterococcus bacteria and Bacteroidesbacteria as described herein to the subject. Following administration ofthe probiotic, communication behavior in the subject can be improved. Insome embodiments, the communication behavior in need of improvement (andsubsequently improved) comprises at least one of communication,sociability, or language comprehension and/or language production. Insome embodiments, the subject is further identified as in need ofimprovement of defects in intestinal barrier integrity, and followingadministration of the probiotic, the defects in defects in intestinalbarrier integrity are improved.

In some embodiments, methods of treating ASD or schizophrenia symptomsare provided. The method can comprise identifying the subject as in needof improving sensorimotor gating and communication behavior. The methodcan comprise administering a probiotic comprising, consistingessentially of, or consisting of an effective amount of Enterococcusbacteria, Bacteroides bacteria, or a combination of Enterococcusbacteria and Bacteroides bacteria as described herein to the subject.Following acute administration of the probiotic, sensorimotor andcommunication behavior in the subject can be improved. In someembodiments, the communication behavior in need of improvement (andsubsequently improved after administration of the probiotic) comprisesat least one of communication, sociability, or language comprehensionand/or language production. In some embodiments, the subject is furtheridentified as in need of improvement of defects in intestinal barrierintegrity, and following administration of the probiotic, the defects indefects in intestinal barrier integrity are improved.

In some embodiments, methods of ameliorating symptoms of ASD orschizophrenia are provided. The method can comprise identifying thesubject as having gastrointestinal or immunological abnormalities orpathologies associated with ASD or schizophrenia. The method cancomprise administering a probiotic comprising, consisting essentiallyof, or consisting of an effective amount of Enterococcus bacteria,Bacteroides bacteria, or a combination of Enterococcus bacteria andBacteroides bacteria as described herein can be administered to thesubject. Following acute administration of the probiotic, the ASD orschizophrenia symptoms can be ameliorated. In some embodiments, thesubject is identified as having ASD or schizophrenia symptoms thatcomprise gastrointestinal abnormalities associated with ASD orschizophrenia, and these symptoms are improved following administrationof the probiotic. In some embodiments, the subject is identified ashaving ASD or schizophrenia symptoms that comprise immunologicalabnormalities associated with ASD or schizophrenia, and these symptomsare improved following administration of the probiotic. In someembodiments, the subject is identified as having ASD or schizophreniasymptoms that comprise immunological and gastrointestinal abnormalitiesassociated with ASD or schizophrenia, and these symptoms are improvedfollowing administration of the probiotic. In some embodiments, thesubject is identified as having ASD symptoms that comprisegastrointestinal abnormalities associated with ASD, and these symptomsare improved following administration of the probiotic. In someembodiments, the subject is identified as having ASD symptoms thatcomprise immunological abnormalities associated with ASD, and thesesymptoms are improved following administration of the probiotic. In someembodiments, the subject is identified as having ASD symptoms thatcomprise immunological and gastrointestinal abnormalities associatedwith ASD, and these symptoms are improved following administration ofthe probiotic. In some embodiments, the subject is identified as havingschizophrenia symptoms that comprise gastrointestinal abnormalitiesassociated with schizophrenia, and these symptoms are improved followingadministration of the probiotic. In some embodiments, the subject isidentified as having schizophrenia symptoms that comprise immunologicalabnormalities associated with schizophrenia, and these symptoms areimproved following administration of the probiotic. In some embodiments,the subject is identified as having schizophrenia symptoms that compriseimmunological and gastrointestinal abnormalities associated withschizophrenia, and these symptoms are improved following administrationof the probiotic. In some embodiments, the subject is further identifiedas in need or improving sensorimotor gating, communication, anxiety,and/or repetitive behavior, and following administration of theprobiotic, the corresponding sensorimotor gating, communication,anxiety, and/or repetitive behavior(s) are improved.

In some embodiments, methods of treating ASD or schizophrenia symptomsare provided. The method can comprise identifying the subject as beingin need of improving repetitive behavior. The method can compriseadministering a probiotic comprising, consisting essentially of, orconsisting of an effective amount of Enterococcus bacteria, Bacteroidesbacteria, or a combination of Enterococcus bacteria and Bacteroidesbacteria as described herein is administered to the subject. Followingacute administration of the probiotic, repetitive behavior in thesubject can be improved.

In some embodiments, methods of preventing ASD or schizophrenia symptomsare provided. The method can comprise identifying the subject as at riskfor developing a sensorimotor gating behavior deficiency. The method cancomprise administering an effective amount of a probiotic comprising,consisting essentially of, or consisting of Enterococcus bacteria,Bacteroides bacteria, or a combination of Enterococcus bacteria andBacteroides bacteria as described herein to the subject in need ofbehavioral improvement. The subject can develop with minimizeddeficiencies or no discernable deficiencies in sensorimotor behavior. Insome embodiments, the ASD or schizophrenia symptoms further comprise atleast one of: impaired communication behavior, anxiety behavior, orrepetitive behavior, and following administration of the probiotic thesubject develops with minimized deficiencies or no discernabledeficiencies in the respective impaired communication behavior, anxietybehavior, or repetitive behavior. In some embodiments, the at-risksubject is an infant or child.

In some embodiments, methods of preventing ASD, or schizophreniasymptoms are provided. The method can comprise identifying a subject asat risk for developing a communication behavior deficiency. The methodcan comprise administering an effective amount of a probioticcomprising, consisting essentially of, or consisting of Enterococcusbacteria, Bacteroides bacteria, or a combination of Enterococcusbacteria and Bacteroides bacteria as described herein is administered tothe subject in need of behavioral improvement. The subject can developwith minimized deficiencies or no discernable deficiencies incommunication behavior. In some embodiments, the communication behaviorin need of improvement (and subsequently improved after administrationof the probiotic) comprises at least one of communication, sociability,or language comprehension and/or language production. In someembodiments, the ASD or schizophrenia symptoms further comprise at leastone of: impaired sensorimotor gating behavior, anxiety behavior, orrepetitive behavior, and following administration of the probiotic thesubject develops with minimized deficiencies or no discernabledeficiencies in sensorimotor gating behavior, anxiety behavior, orrepetitive behavior is improved. In some embodiments, the at-risksubject is an infant or child.

In some embodiments, methods of preventing ASD symptoms are provided.The method can comprise identifying a subject as at risk for developinga communication behavior deficiency and sensory gating behaviordeficiency. The method can comprise administering an effective amount ofa probiotic comprising, consisting essentially of, or consisting ofEnterococcus bacteria, Bacteroides bacteria, or a combination ofEnterococcus bacteria and Bacteroides bacteria as described herein isadministered to the subject at risk. The subject can develop withminimized deficiencies or no discernable deficiencies in socialcommunication behavior. In some embodiments, the communication behaviorcomprises at least one of communication, sociability, or languagecomprehension and/or language production. Optionally, the at-risksubject is an infant or child.

In some embodiments, for any of the above methods, the probioticcomprising, consisting essentially of, or consisting of Enterococcusbacteria, Bacteroides bacteria, or a combination of Enterococcusbacteria and Bacteroides bacteria of any of the methods described hereinis selected from the group consisting of: (a) Enterococcus bacteria(e.g., E. faecilis, E. faecium, E. hirae, E. avium, E. durans, E.gallinarum, or E. casseliflavus); (b) Enterococcus bacteria (e.g., E.faecilis, E. faecium, E. hirae, E. avium, E. durans, E. gallinarum, orE. casseliflavus); and Bacteroides bacteria (e.g., B. fragilis, B.thetaiotaomicron or B. vulgatus); (c) Enterococcus bacteria and B.fragilis; (d) Enterococcus bacteria and B. thetaiotaomicron; (d)Enterococcus bacteria and B. vulgatus; (e) Enterococcus bacteria, B.fragilis, and B. thetaiotaomicron, (f) Enterococcus bacteria, B.fragilis, and B. vulgatus; (g) Enterococcus bacteria, B.thetaiotaomicron and B. vulgatus; (h) Enterococcus bacteria, B.fragilis, B. thetaiotaomicron and B. vulgatus; (i) E. faecilis and B.fragilis; (j) E. faecilis and B. thetaiotaomicron; (k) E. faecilis andB. vulgatus; (l) E. faecilis, B. fragilis, and B. thetaiotaomicron, (m)E. faecilis, B. fragilis, and B. vulgatus; (n) E. faecilis, B.thetaiotaomicron and B. vulgatus; (o) E. faecilis, B. fragilis, B.thetaiotaomicron and B. vulgatus; (p) Bacteroides bacteria; (q) B.fragilis and B. thetaiotaomicron; (r) B. fragilis, and B. vulgatus; (s)B. thetaiotaomicron and B. vulgatus; or (t) B. fragilis, B.thetaiotaomicron and B. vulgatus. Optionally, the subject isadministered no other bacteria, or substantially no other bacteria apartfrom the identified bacteria of the probiotic, and as such the probioticfor use in treatment of the subject is in a composition or compositionsfree or substantially free of other bacteria. Optionally, the subject isadministered no antibiotics, or is administered substantially noantibiotics, and as such the probiotic for administration to the subjectis in a composition or compositions free or substantially free ofantibiotics. Optionally, the subject is administered no drugs, or isadministered substantially no drugs, and as such the probiotic foradministration to the subject is in a composition or compositions freeor substantially free of drugs. Optionally, the subject is administeredno pharmaceutically active ingredients, or is administered substantiallyno pharmaceutically active ingredients, and as such the probiotic foradministration to the subject is in a composition or compositions freeor substantially free of pharmaceutically active ingredients.

In some embodiments as described above, the method further comprisesdetermining that the subject is in need of improving a behavior. In someembodiments, for example uses, methods, and/or compositions directed toinfants and/or children, a subject at risk for an ASD behavior isidentified based on maternal immune activation and/or other riskfactors. In some embodiments, the subject is diagnosed as having ASDbased on the level of an ASD-related metabolite or combination ofmetabolites in the gut, in a bodily fluid (for example, blood andurine), or any combination thereof. Methods of diagnosing ASD based onlevels of metabolite in a subject are described in detail in US Pub. No.2014/0065132, hereby incorporated by reference in its entirety. In someembodiments, the subject is determined to have a lesion or developmentaldeficiency in a region of the brain associated with speech production,speech recognition, impulse control, and socialization, for exampleregions of the cerebral cortex, the corpus callosum, Broca's area,and/or Wernicke's area. In some embodiments, an ASD behavior, forexample a deficient communication, vocalization, sensorimotor, anxiety,and/or repetitive behavior, or a combination of two or more of these isidentified using standard diagnostic criteria, for example in theDiagnostic and Statistical Manual of Mental Disorders, Fourth Edition(DSM-4) or Fifth Edition (DSM-5). In some embodiments, the presence orabsence of ASD in the subject is determined using a behavioral test, forexample at least one of the Autism Behavior Checklist (ABC), Autismdiagnostic Interview-Revised (ADI-R), childhood autism Rating Scale(CARS), and/or Pre-Linguistic Autism Diagnostic Observation Schedule(PL-ADOS). The behavioral test can include, but is not limited to,detecting the presence and/or extent of 1) preoccupation with one ormore stereotyped and restricted patterns of interest that is abnormal ineither intensity or focus, 2) inflexible adherence to specific,nonfunctional routines or rituals, c) stereotyped and repetitive motormannerisms (such as hand flapping, finger flapping etc.), and/or d)persistent preoccupation with parts of objects. Non-limiting examples ofbehavior that can be included in a behavioral test and suggest a needfor improving behavioral performance in the subject under the testinclude: a) sensory behaviors, including poor use of visualdiscrimination when learning, seems not to hear, so that a hearing lossis suspected, sometimes shows no “startle response” to loud noise”,sometimes painful stimuli such as bruises, cuts, and injections evoke noreaction, often will not blink when bright light is directed towardeyes, covers ears at many sounds, squints, frowns, or covers eyes whenin the presence of natural light, frequently has no visual reaction to a“new” person, stares into space for long periods of time; b) relatingbehaviors: frequently does not attend to social/environmental stimuli,has no social smile, does not reach out when reached for, non-responsiveto other people's facial expressions/feelings, actively avoids eyecontact, resists being touched or held, is flaccid when held in arms, isstiff and hard to held, does not imitate other children at play, has notdeveloped any friendships, often frightened or very anxious, “looksthrough” people; c) body and object use behaviors: whirls self for longperiods of time, does not use toys appropriately, insists on keepingcertain objects with him/her, rocks self for long periods of time, doesa lot of lunging and darting, flaps hands, walks on toes, hurts self bybanging head, biting hand, twirls, spins, and bangs objects a lot, feel,smell, and/or taste objects in the environment, gets involved incomplicated “rituals” such as lining things up, is very destructive; andd) language behaviors: does not follow simple commands given once, haspronoun reversal, speech is atonal, does not respond to own name whencalled out among two others, seldom says “yes” or “I”, does not followsimple commands involving prepositions, gets desired objects bygesturing, repeats phrases over and over, cannot point to more than fivenamed objects, uses 0-5 spontaneous words per day to communicate wantsand needs, repeats sounds or words over and over, echoes questions orstatements made by others, uses at least 15 but less than 30 spontaneousphrases daily to communicate, learns a simple task but “forgets”quickly, strong reactions to changes in routine/environment, has“special abilities” in one area of development, which seems to rule outmental retardation, severe temper tantrums and/or frequent minortantrums, hurts others by biting, hitting, and/or kicking, does not waitfor needs to be met, difficulties with toileting, does not dress selfwithout frequent help, frequently unaware of surroundings, and may beoblivious to dangerous situations, prefers to manipulate and be occupiedwith inanimate things, and/or a developmental delay identified at orbefore 30 months of age. One of ordinary skill in the art wouldappreciate that the attending physician would know how to identify asubject in need of treatment disclosed herein.

In some embodiments as described above, the method comprisesadministering the effective amount of probiotic in a singleadministration of one or more compositions. In some embodiments asdescribed above, the method comprises administering the effective amountof the probiotic across two or more administrations of a singlecomposition as described herein. For example, the compositions can beadministered about 1 minute, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30,35, 40, 45, 50, 55 minutes, 1 hour, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 hours, 1 day, 2, 3, 4, 5, 6,7, 8, 9, or 10 days apart, including ranges between any two of thelisted values, for example 1 minute-10 minutes, 1 minute to 30 minutes,1 minute to 1 hour, 1 minute-2 hours, 1 minute-4 hours, 1 minute-12hours, 1 minute-18 hours, 1 minute-1 day, 10 minutes to 30 minutes, 10minutes to 1 hour, 10 minutes-2 hours, 10 minutes-4 hours, 10 minute-12hours, 10 minutes-18 hours, 10 minutes-1 day, 30 minutes to 1 hour, 30minutes-2 hours, 30 minutes-4 hours, 30 minute-12 hours, 30 minutes-18hours, 30 minutes-1 day, 30 minutes-2 days, 1 hour-2 hours, 1 hour-4hours, 1 hour-12 hours, 1 hour-18 hours, 1 hour-1 day, 4 hours-12 hours,4 hours-18 hours, 4 hours-1 day, 1 day-2 days, 1 day-3 days, 1 day-4days, 1 day-5 days, 1 day-7 days, 1 day-10 days, 2 days-3 days, 2 days-4days, 2 days-5 days, 2 days-7 days, 2 days-10 days, or 5 days to 10days. In some embodiments as described above, the method comprisesadministering the effective amount of two or more different compositionsas described herein across two or more administrations of a singlecomposition. For example, the second composition can be administeredabout 1 minute, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45,50, 55 minutes, 1 hour, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, 22, 23 hours, 1 day, 2, 3, 4, 5, 6, 7, 8, 9, or10 days after the first composition, including ranges between any two ofthe listed values, for example 1 minute-10 minutes, 1 minute to 30minutes, 1 minute to 1 hour, 1 minute-2 hours, 1 minute-4 hours, 1minute-12 hours, 1 minute-18 hours, 1 minute-1 day, 10 minutes to 30minutes, 10 minutes to 1 hour, 10 minutes-2 hours, 10 minutes-4 hours,10 minute-12 hours, 10 minutes-18 hours, 10 minutes-1 day, 30 minutes to1 hour, 30 minutes-2 hours, 30 minutes-4 hours, 30 minute-12 hours, 30minutes-18 hours, 30 minutes-1 day, 30 minutes-2 days, 1 hour-2 hours, 1hour-4 hours, 1 hour-12 hours, 1 hour-18 hours, 1 hour-1 day, 4 hours-12hours, 4 hours-18 hours, 4 hours-1 day, 1 day-2 days, 1 day-3 days, 1day-4 days, 1 day-5 days, 1 day-7 days, 1 day-10 days, 2 days-3 days, 2days-4 days, 2 days-5 days, 2 days-7 days, 2 days-10 days, or 5 days to10 days. In some embodiments, the probiotic is administered in aslow-release formulation (for example a slow-release capsule or implant)for any of the durations described above.

In some embodiments, the probiotic is administered to the subject untilan improvement in behavioral performance is observed. Optionally, theprobiotic is administered to the subject after an improvement inbehavioral performance is observed, for example to solidify or maintainthe improved behavioral performance.

EXAMPLES

Some aspects of the embodiments discussed above are disclosed in furtherdetail in the following examples, which are not in any way intended tolimit the scope of the present disclosure.

Example 1: Acute Postnatal Treatment with B. fragilis, dPSA B. fragilis,and B. vulgatus, but not E. faecalis Corrects Defects in IntestinalBarrier Integrity in the MIA Mouse Model for Autism

Offspring of immune-activated mothers were given applesauce containingeach bacterium or applesauce alone for one week post-weaning. Six weeksafter bacterial treatment, mice were orally gavaged with FITC-dextran tomeasure intestinal permeability in vivo, alongside a positive control ora chemically-induced colitis mouse model. Poly(I:C) was administered asan immune activator in mothers. The results are shown in FIG. 1.Treatment with B. fragilis (n=9), B. thetaiotaomicron (n=7), and B.vulgatus (n=5) significantly reduced leakage of FITC-dextran into thebloodstream in the MIA mouse model for autism. E. faecalis (n=6)treatment had no significant effect.

Thus, acute postnatal treatment with B. fragilis, mutant B. fragilislacking polysaccharide A (dPSA), B. thetaiotaomicron, and B. vulgatus,but not E. faecalis corrects defects in intestinal barrier integrity inthe MIA mouse model for autism in accordance with some embodimentsdescribed herein.

Example 2: Acute Postnatal Treatment with B. fragilis, dPSA B. fragilis,B. thetaiotaomicron, and B. vulgatus, but not E. faecalis CorrectsAnxiety-Like Behaviors in the MIA Mouse Model for Autism

Offspring of immune-activated mothers were given applesauce containingeach bacterium or applesauce alone for 1 week post-weaning. Poly(I:C)was administered as an immune activator in mothers. 3-6 weeks afterbacterial treatment, mice were tested for anxiety-like and exploratorybehavior in the open field exploration task. Treatment with B. fragilis(n=47), B. thetaiotaomicron (n=32), and B. vulgatus (n=20) reducedanxiety-like behavior in the MIA mouse model for autism, as measured byincreased entries into the center of the open area (FIG. 2A), andincreased duration spent in the center of the open area (FIG. 2B). E.faecalis (n=16) treatment had no significant effect in either of theseassays.

Thus, acute postnatal treatment with B. fragilis, mutant B. fragilislacking polysaccharide A (dPSA), B. thetaiotaomicron, and B. vulgatus,but not E. faecalis corrects anxiety-like behaviors in the MIA mousemodel for autism in accordance with some embodiments described herein.

Example 3: Acute Postnatal Treatment with B. fragilis, dPSA B. fragilis,B. thetaiotaomicron, and B. vulgatus, but not E. faecalis ImprovesRepetitive Behaviors in the MIA Mouse Model for Autism

Offspring of immune-activated mothers were given applesauce containingeach bacterium or applesauce alone for 1 week post-weaning. 3-6 weeksafter bacterial treatment, mice were tested for repetitive behavior inthe stereotyped marble burying task. Treatment with B. fragilis (n=30),B. thetaiotaomicron (n=32), and B. vulgatus (n=18) reduced compulsivemarble burying in the MIA mouse model for autism. E faecalis (n=16) hadno apparent effect in this assay.

Thus, acute postnatal treatment with B. fragilis, mutant B. fragilislacking polysaccharide A (dPSA), B. thetaiotaomicron, and B. vulgatus,but not E. faecalis improves repetitive behaviors in the maternal immuneactivation (MIA) mouse model for autism in accordance with someembodiments described herein.

Example 4: Acute Postnatal Treatment with B. fragilis, B.thetaiotaomicron, B. vulgatus, and E. faecalis Improves CommunicativeBehavior in the MIA Mouse Model for Autism

Offspring of immune-activated mothers were given applesauce containingeach bacterium or applesauce alone for 1 week post-weaning. 3-6 weeksafter bacterial treatment, mice were tested for ultrasonic vocalizationsin response to exposure to unfamiliar female stimulus mouse. Treatmentwith B. fragilis (n=10), B. thetaiotaomicron (n=10), and B. vulgatus(n=10) and E. faecalis (n=10) significantly increased the total numberof calls produced (see FIG. 4A), significantly increased averageduration per call (see FIG. 4B), and significantly increased totalduration of vocalization (see FIG. 4C) in the MIA mouse model of autism.Mutant B. fragilis lacking polysaccharide A (dPSA) treatment has nosignificant effect.

Thus, acute postnatal treatment with B. fragilis, B. thetaiotaomicron,B. vulgatus, and E. faecalis improves communicative behavior (e.g. totalnumber of calls, average call duration, and total call duration) in theMIA mouse model in accordance with some embodiments described herein.

Example 5: Acute Postnatal Treatment with B. fragilis, B.thetaiotaomicron, and E. faecalis Improves Sensorimotor Behavior in theMIA Mouse Model for Autism

Offspring of immune-activated mothers were given applesauce containingeach bacterium or applesauce alone for 1 week post-weaning. 3-6 weeksafter bacterial treatment, mice were tested for sensorimotor behavior inthe pre-pulse inhibition task. As shown in FIG. 5, Treatment with B.fragilis (n=40), B. thetaiotaomicron (n=32), and E. faecalis (n=16)elevate pre-pulse inhibition in response to a 5 decibel (PP15) and 15decibel (PPI15) pre-pulse. B. vulgatus (n=20) treatment has no apparenteffect in this assay.

As shown in FIG. 5, statistically significant differences are observedfor a number of bacteria in accordance with some embodiments describedherein, as summarized in Table 5. Sensorimotor behavior as measured by %PPI was compared for various treatments. In Table 5, p<0.05 is denotedwith a single asterisk (*), and p<0.01 is denoted with a double asterisk(**).

TABLE 5 Saline vs saline B. frag: 0.0059** Saline vs polyIC 0.0100*Saline vs polyIC B. frag: 0.92776 Saline vs polyIC B frag dPSA 0.1074Saline vs polyIC B. vulgatus 0.1415 polyIC vs polyIC B. frag 0.0189polyIC vs polyIC B. frag dPSA 0.8173 polyIC vs polyIC B. theta 0.0062**polyIC vs polyIC B. vulgatus 0.6763 polyIC vs polyIC E. faecalis 0.0712

Thus, acute postnatal treatment with B. fragilis, B. thetaiotaomicron,and E. faecalis improves sensorimotor behavior in the MIA mouse modelfor autism in accordance with some embodiments described herein.

Example 6: Treatment with E. faecalis Improves Sensorimotor Behavior ina Human Exhibiting Sensorimotor Deficiencies

A human child subject is identified as having ASD based on the childhoodautism Rating Scale (CARS), and exhibits limited sensorimotor responseto auditory stimuli. The subject drinks a yogurt comprising an effectiveamount of a probiotic consisting essentially of E. faecalis weekly forthree weeks. After about three weeks of drinking the yogurt, the subjectis expected to exhibit increased response to auditory stimuli.

Example 7: Treatment with E. faecalis Improves Vocalization andCommunication Behavior in a Human Exhibiting Vocalization andCommunication Deficiencies

A human ASD adult subject exhibits limited verbalization andcommunication behavior in response to verbal prompts from others. Thesubject swallows a gel capsule comprising an effective amount of aprobiotic comprising E. faecalis and substantially free of otherbacteria and antibiotics daily until the subject exhibits increasedverbalization and communication behavior in response to verbal promptsfrom others.

Example 8: Treatment with B. thetaiotaomicron and E. faecalis ImprovesCommunication Behavior and Anxiety Behavior in a Human ExhibitingCommunication and Anxiety Deficiencies

A human ASD adolescent subject exhibits deficient communication andanxiety behaviors. The subject is fed a first yogurt beverage comprisingan effective amount of B. thetaiotaomicron and a second yogurt beveragecomprising an effective amount of E. faecalis once every four days. Theadministration continues on this schedule for eight weeks. After eightweeks, the subject is expected to exhibit improved communication andanxiety behaviors.

Example 9: Acute Postnatal Treatment with E. faecalis ImprovesSensorimotor and Communication Behavior and Anxiety Behavior in a HumanExhibiting Communication and Anxiety Deficiencies

A human infant subject at risk for ASD sensorimotor and communicationbehavior deficiencies is fed a composition comprising applesauce and aneffective amount of E. faecalis in a weekly doses post-weaning for tenweeks. The subject is expected to develop without ASD-like sensorimotorand communication behaviors.

In at least some of the previously described embodiments, one or moreelements used in an embodiment can interchangeably be used in anotherembodiment unless such a replacement is not technically feasible. Itwill be appreciated by those skilled in the art that various otheromissions, additions, and modifications may be made to the methods andstructures described above without departing from the scope of theclaimed subject matter. All such modifications and changes are intendedto fall within the scope of the subject matter, as defined by theappended claims.

With respect to the use of substantially any plural and/or singularterms herein, those having skill in the art can translate from theplural to the singular and/or from the singular to the plural as isappropriate to the context and/or application. The varioussingular/plural permutations may be expressly set forth herein for sakeof clarity.

It will be understood by those within the art that, in general, termsused herein, and especially in the appended claims (e.g., bodies of theappended claims) are generally intended as “open” terms (e.g., the term“including” should be interpreted as “including but not limited to,” theterm “having” should be interpreted as “having at least,” the term“includes” should be interpreted as “includes but is not limited to,”etc.). It will be further understood by those within the art that if aspecific number of an introduced claim recitation is intended, such anintent will be explicitly recited in the claim, and in the absence ofsuch recitation no such intent is present. For example, as an aid tounderstanding, the following appended claims may contain usage of theintroductory phrases “at least one” and “one or more” to introduce claimrecitations. However, the use of such phrases should not be construed toimply that the introduction of a claim recitation by the indefinitearticles “a” or “an” limits any particular claim containing suchintroduced claim recitation to embodiments containing only one suchrecitation, even when the same claim includes the introductory phrases“one or more” or “at least one” and indefinite articles such as “a” or“an” (e.g., “a” and/or “an” should be interpreted to mean “at least one”or “one or more”); the same holds true for the use of definite articlesused to introduce claim recitations. In addition, even if a specificnumber of an introduced claim recitation is explicitly recited, thoseskilled in the art will recognize that such recitation should beinterpreted to mean at least the recited number (e.g., the barerecitation of “two recitations,” without other modifiers, means at leasttwo recitations, or two or more recitations). Furthermore, in thoseinstances where a convention analogous to “at least one of A, B, and C,etc.” is used, in general such a construction is intended in the senseone having skill in the art would understand the convention (e.g., “asystem having at least one of A, B, and C” would include but not belimited to systems that have A alone, B alone, C alone, A and Btogether, A and C together, B and C together, and/or A, B, and Ctogether, etc.). In those instances where a convention analogous to “atleast one of A, B, or C, etc.” is used, in general such a constructionis intended in the sense one having skill in the art would understandthe convention (e.g., “a system having at least one of A, B, or C” wouldinclude but not be limited to systems that have A alone, B alone, Calone, A and B together, A and C together, B and C together, and/or A,B, and C together, etc.). It will be further understood by those withinthe art that virtually any disjunctive word and/or phrase presenting twoor more alternative terms, whether in the description, claims, ordrawings, should be understood to contemplate the possibilities ofincluding one of the terms, either of the terms, or both terms. Forexample, the phrase “A or B” will be understood to include thepossibilities of “A” or “B” or “A and B.”

In addition, where features or aspects of the disclosure are describedin terms of Markush groups, those skilled in the art will recognize thatthe disclosure is also thereby described in terms of any individualmember or subgroup of members of the Markush group.

As will be understood by one of skill in the art, for any and allpurposes, such as in terms of providing a written description, allranges disclosed herein also encompass any and all possible sub-rangesand combinations of sub-ranges thereof. Any listed range can be easilyrecognized as sufficiently describing and enabling the same range beingbroken down into at least equal halves, thirds, quarters, fifths,tenths, etc. As a non-limiting example, each range discussed herein canbe readily broken down into a lower third, middle third and upper third,etc. As will also be understood by one skilled in the art all languagesuch as “up to,” “at least,” “greater than,” “less than,” and the likeinclude the number recited and refer to ranges which can be subsequentlybroken down into sub-ranges as discussed above. Finally, as will beunderstood by one skilled in the art, a range includes each individualmember. Thus, for example, a group having 1-3 articles refers to groupshaving 1, 2, or 3 articles. Similarly, a group having 1-5 articlesrefers to groups having 1, 2, 3, 4, or 5 articles, and so forth.

While various aspects and embodiments have been disclosed herein, otheraspects and embodiments will be apparent to those of skill in the art.The various aspects and embodiments disclosed herein are for purposes ofillustration and are not intended to be limiting, with the true scopeand spirit being indicated by the following claims.

What is claimed is:
 1. A composition comprising: Enterococcus bacteria;and Bacteroides bacteria.
 2. The composition of claim 1, wherein thecomposition is substantially free of bacteria other than E. faecalis, B.fragilis, B. thetaiotaomicron, and B. vulgatus.
 3. The composition ofclaim 1, wherein the composition is substantially free of bacteria otherthan E. faecalis, B. thetaiotaomicron, and B. vulgatus.
 4. Thecomposition of claim 1, wherein the Enterococcus bacteria comprises E.faecilis.
 5. The composition of claim 1, wherein a sole activeingredient consists essentially of: a mixture of Enterococcus bacteriaand B. fragilis; a mixture of Enterococcus bacteria and B.thetaiotaomicron; a mixture of Enterococcus bacteria and B. vulgatus; amixture of Enterococcus bacteria, B. fragilis, and B. thetaiotaomicron:a mixture of Enterococcus bacteria, B. fragilis, and B. vulgatus; amixture of Enterococcus bacteria, B. thetaiotaomicron, and B. vulgatus:or a mixture of Enterococcus bacteria, B. fragilis, B. thetaiotaomicron,and B. vulgatus.
 6. The composition of claim 1, wherein the Enterococcusbacteria comprises E. faecalis.
 7. The composition of claim 1, whereinthe Enterococcus bacteria comprises at least about 10⁷ colony formingunits (cfu).
 8. The composition of claim 1, wherein the composition isformulated as a food composition.
 9. The composition of claim 8, whereinthe food composition comprises milk, yogurt, curd, cheese, fermentedmilks, milk based fermented products, ice creams, fermented cereal-basedproducts, milk-based powders, infant formula, tablets, liquid bacterialsuspensions, dried oral supplement, or wet oral supplement.
 10. Thecomposition of claim 1, wherein the composition is formulated as aprobiotic composition.
 11. The composition of claim 1, wherein thecomposition is formulated as a dietary supplement.
 12. The compositionof claim 11, wherein the dietary supplement is a capsule, tablet,powder, or liquid.